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>   home   >   Products   >   Primary Antibodies   >   Anti-V5-Tag Antibody, clone SV5-Pk1 (DyLight®405)   

Anti-V5-Tag Antibody, clone SV5-Pk1

Mouse Anti-Viral Monoclonal Antibody

     
  • WB - Anti-V5-Tag Antibody, clone SV5-Pk1  ABD12063
    Published customer image:Mouse anti V5 tag antibody, clone SV5-Pk1 used for detection of V5 tagged REDD1 protein by western blotting.Image caption:REDD1 is not degraded by Cul4a or phosphorylated by GSK3ß at Thr 23 and Thr25. (A) HEK293 cells were transfected with empty vector or tetracycline inducible dnCul4a-V5 pcDNA4/TO (1 µg) and Cul4A-V5 pcDNA3 (0.03 µg) followed by tetracycline (1 µg/ml) induction for 24 hours and cell lysis. (B) HEK293 cells were transfected with 0.5 µg Cul4a-V5 pcDNA3 for 15 hours followed by transfection of 20 nM control or Cul4a siRNAs to determine siRNAs efficiency. (C) HEK293 cells were transfected with 20 nM control or Cul4a siRNAs for 3 days followed by cell lysis. (D) REDD1-V5 pcDNA3 wild type, T23A T25A or T23D T25D plasmids (0.4 µg) were transfected in HEK293 cells for 3 days and treated with 20 µM MG-132 for 6 hours followed by cell lysis. (E) HEK293 cells were treated with 30 mM LiCl or GSK3 inhibitor IX (5 µM or 10 µM) for 20 hours followed by cell lysis. (F) HEK293 cells were co-transfected with 0.2 µg REDD1-V5 pcDNA3 and 0.3 µg GSK3ß pcDNA3 or empty pcDNA3 for 3 days followed by MG-132 (20 µM) treatment for 6 hours followed by cell lysis. (G) HEK293 cells were transfected with 3 µg FLAG-REDD1 or FLAG-FRAT1 for 3 days followed by cell lysis and FLAG immunoprecipitation. In vitro phosphorylation of REDD1 and FRAT1 was carried out as described in Materials and Methods..From: Tan CY, Hagen T mTORC1 Dependent Regulation of REDD1 Protein Stability. PLoS ONE 8: e63970.
  • WB - Anti-V5-Tag Antibody, clone SV5-Pk1  ABD12063
    Published customer image:Mouse anti V5 Tag antibody, clone SV5-Pk1used for immunoprecipitation of V5 tagged RNF11proteinImage caption:Myristoylation mutant of RNF11 is unable to associate with Itch. SH-SY5Y shRNA-RNF11 cells were transfected with shRNA-resistant RNF11 constructs or vector. Coimmunoprecipitation experiments using V5 antibody were performed 24 hours after transfection. Immunoprecipitates and lysates were resolved by SDS-PAGE and immunoblotted with anti-A20, Itch, RNF11 or actin. Blots are representative of three independent experiments. From: Pranski EL, Dalal NV, Herskowitz JH, Orr AL, Roesch LA, Fritz JJ, Heilman C, Lah JJ, Levey AI, Betarbet RS. Neuronal RING finger protein 11 regulates canonical NF-?B signaling. J Neuroinflammation. 2012 Apr 16;9:67. doi: 10.1186/1742-2094-9-67.
  • WB - Anti-V5-Tag Antibody, clone SV5-Pk1  ABD12063
    Published customer image:Mouse anti V5 tag antibody, clone SV5-Pk1 used for the detection of V5 tagged HIVA peptide by western blottingImage caption:Construction of the BCG.HIVACAT vaccine strain. (A) A synthetic GC-rich HIVA gene was fused to the region encoding the 19-kDa lipoprotein signal sequence and inserted into the episomal pJH222 E. coli-mycobacterium shuttle plasmid. This plasmid contains kanamycin resistance and complementing lysA genes and an E. coli origin of replication . In addition, pJH222 contained the mycobacterial origin of replication . The BALB/c mouse T-cell and MAb Pk epitopes used in this work are depicted. P a-Ag, M. tuberculosis a-antigen promoter; PHSP60, heat shock protein 60 gene promoter. The aph gene was removed by SpeI digestion and the lacO sequence was inserted and transformed into E. coli DH1lacdapD strain. (B) Immunodot of BCG.HIVACAT lysates. Dot 1: BCG wild type (negative control); Dot 2, 3, 4 and 5: clone 3, clone 7, clone 9 and clone 10 of BCG.HIVACAT; Dot 6: BCG.HIVA222 (positive control). HIVA peptide was detected using the anti-Pk MAb followed by horseradish peroxidase-Goat-anti-Mouse and enhanced chemiluminescence detection. (C) In vivo plasmid stability of BCG.HIVACAT harboring pJH222.HIVACAT. Mice were injected s.c. with 105 cfu of BCG.HIVACAT and boosted i.m. with 106 pfu of MVA.HIVA, spleens were homogenized 20 weeks after BCG inoculation and the recovered rBCG colonies were tested for the presence of the HIVA DNA coding sequence by PCR. Lanes 1 to 6: Six rBCG colonies were recovered in the non-lysine supplemented plate; lane 7: Molecular weight marker; lane 8: Plasmid DNA positive control; lane 9: Distilled water (negative control).From: Saubi N, Mbewe-Mvula A, Gea-Mallorqui E, Rosario M, Gatell JM, et al. Pre-Clinical Development of BCG.HIVACAT, an Antibiotic-Free Selection Strain, for HIV-TB Pediatric Vaccine Vectored by Lysine Auxotroph of BCG. PLoS ONE 7(8): e42559.
  • WB - Anti-V5-Tag Antibody, clone SV5-Pk1  ABD12063
    Published customer image:Mouse anti V5 tag antibody, clone SV5-Pk1 used for detection of V5 tagged REDD1 protein by western blotting.Image caption:REDD1 is not regulated by Cullin E3 Ubiquitin ligases. (A,B) Untransfected (A) or REDD1-V5 pcDNA3 (0.3 µg) transfected (B) HEK293 cells stably expressing tetracycline inducible dnUbc12-HA were induced with 1 µg/ml tetracycline for 24 hours (A,B) or treated with 3 µM MLN4924 (A) or 20 µM MG-132 (A,B) for 8 hours followed by cell lysis. (C,D) Untransfected HEK293 (C) or HEK293 transfected with REDD1-V5 pcDNA3 (0.3 µg) (D) were pre-treated with 3 µM MLN4924 followed by cycloheximide (40 µM) treatment and cell lysis at the indicated time points. (E) HEK293 cells stably expressing tetracycline inducible dnUbc12-HA were induced with 1 µg/ml tetracycline for 24 hours followed by cycloheximide (40 µM) treatment and cell lysis at the indicated time points.From: Tan CY, Hagen T mTORC1 Dependent Regulation of REDD1 Protein Stability. PLoS ONE 8(5): e63970.
  • WB - Anti-V5-Tag Antibody, clone SV5-Pk1  ABD12063
    Published customer image:Mouse anti V5 tag antibody, clone SV5-Pk1 used for detection of V5 tagged podocin protein by western blottingImage caption:The podocin short isoform interacts with known podocyte proteins. CD2AP, TRPC6, neprin and NEPH1 co-precipitate with both podocin isoforms (FL, full length (canoncical isoform); SI, short isoform). FLAG- and V5-tagged proteins were expressed in HEK293T cells and precipitated with anti-FLAG antibody as indicated. Western blot analysis was performed with a V5 specific antibody. Expression levels of FLAG.podocin constructs in the lysates are shown below.From: Völker LA, Schurek EM, Rinschen MM, Tax J, Schutte BA, Lamkemeyer T, Ungrue D, Schermer B, Benzing T, Höhne M. Characterization of a short isoform of the kidney protein podocin in human kidney. BMC Nephrol. 2013 May 6;14:102.
  • SPECIFICATION
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Product Information
Application
  • Applications Legend:
  • WB=Western Blot
  • IHC=Immunohistochemistry
  • IHC-P=Immunohistochemistry (Paraffin-embedded Sections)
  • IHC-F=Immunohistochemistry (Frozen Sections)
  • IF=Immunofluorescence
  • FC=Flow Cytopmetry
  • IC=Immunochemistry
  • ICC=Immunocytochemistry
  • E=ELISA
  • IP=Immunoprecipitation
  • DB=Dot Blot
  • CHIP=Chromatin Immunoprecipitation
  • FA=Fluorescence Assay
  • IEM=Immunoelectronmicroscopy
  • EIA=Enzyme Immunoassay
WB, IHC-F, IF, FC, E, IP
Primary Accession P11207
Reactivity Virus
Host Mouse
Clonality Monoclonal
Isotype IgG2a
Clone Names SV5-Pk1
Calculated MW 23935 Da
Additional Information
Purification Purified IgG prepared by affinity chromatography on Protein G from tissue culture supernatant
Immunogen Paramyxovirus Simian-Virus 5 (SV5)
Shelf Life 18 months from date of despatch.
Gene ID 3160800
Other Names Non-structural protein V, P/V
Target/Specificity Mouse anti-V5-Tag, clone SV5-Pk1 recognizes the sequence, IPNPLLGL, present on the P/V proteins of the paramyxovirus, SV5 (Dunnet al.1999). Clone SV5-Pk1 is used to detect recombinant proteins, some of which include transmembrane and secreted proteins, that have labelled with tags containing this sequence (Randallet al.1993andZhaoet al.2005).
Preservative & Stabilisers 0.09% Sodium Azide
Storage Store at +4℃ or at -20 ℃.
PrecautionsAnti-V5-Tag Antibody, clone SV5-Pk1 is for research use only and not for use in diagnostic or therapeutic procedures.
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References

1. Southern, J.A. et al. (1991) Identification of an epitope on the P and V proteins of simian virus 5 that distinguishes between two isolates with different biological characteristics.
J. Gen. Virol. 72: 1551-1557. 2. Orime, K. et al. (2013) Trefoil Factor 2 Promotes Cell Proliferation in Pancreatic ß-Cells through CXCR-4-Mediated ERK1/2 Phosphorylation.
Endocrinology. 154: 54-64. 3. Randall, R.E. et al. (1993) Two-tag purification of recombinant proteins for the construction of solid matrix-antibody antigen (SMAA) complexes as vaccines.
Vaccine 11:1247-1252. 4. Randall, R.E. et al. (1994) Purification of antibody-antigen complexes containing recombinant SIV proteins; comparison of antigen and antibody-antigen complexes for immune priming.
Vaccine 12: 351-358. 5. Hanke, T. et al. (1995) Attachment of an oligopeptide epitope to the C-terminus of recombinant SIV gp160 facilitates the construction of SMAA complexes while preserving CD4 binding
J. Virol. Methods 149-156. 6. Jaffray, E. et al. (1995) Domain organization of I kappa B alpha and sites of interaction with NF-kappa B p65
Mol. Cell. Biol. 15: 2166-2172. 7. Rodriguez, M.S. et al. (1995) Inducible degradation of I kappa B alpha in vitroandin vivo requires the acidic C-terminal domain of the protein.
Mol. Cell. Biol. 15: 2413-2419. 8. Arenzana-Seisdedos, F. et al. (1995) Inducible nuclear expression of newly synthesised IkBa negatively regulates DNA binding and transcriptional activities of NF-kB.
Mol. Cell. Biol. 15: 2689-2696. 9. Hirst, K. et al. (1994) The transcription factor, the Cdk, its cyclin and their regulator: directing the transcriptional response to a nutritional signal
EMBO J. 13: 5410-5420. 10. Dunn, C. et al. (1999) Fine mapping of the binding sites of monoclonal antibodies raised against the Pk tag.
J. Immunol. Methods. 224: 141-150. 11. Young, D.F. et al. (2001) Single amino acid substitution in the V protein of simian virus 5 differentiates its ability to block interferon signaling in human and murine cells
J. Virol. 75 (7): 3363 - 3370. 12. Stuart A. MacNeill (2004) The C-terminal zinc finger of the catalytic subunit of DNA polymerase {delta} is responsible for direct interaction with the B-subunit
Nucleic Acids Research 32: 3005-3016 13. Herskowitz, J.H. et al. (2011) Rho Kinase II Phosphorylation of the Lipoprotein Receptor LR11/SORLA AltersAmyloid-{beta} Production.
J Biol Chem. 2011 286: 6117-27. 14. Liebau, M.C. et al. (2011) Nephrocystin-4 regulates Pyk2-induced tyrosine phosphorylation of Nephrocystin-1 to control targeting to monocilia.
J Biol Chem. 286: 14237-45. 15. Björk, J.K. et al. (2010) miR-18, a member of Oncomir-1, targets heat shock transcription factor 2 in spermatogenesis.
Development. 2010 Oct;137(19):3177-84. 16. Boggio, R. et al. (2007) Targeting SUMO E1 to ubiquitin ligases: a viral strategy to counteract sumoylation.
J Biol Chem. 282: 15376-82. 17. Gallazzini, M. et al. (2011) High NaCl-induced activation of CDK5 increases phosphorylation of the osmoprotective transcription factor TonEBP/OREBP at threonine 135, which contributes to its rapid nuclear localization.
Mol Biol Cell. 22: 703-14. 18. Hadler, K.S. et al. (2008) Identification of a non-purple tartrate-resistant acid phosphatase: an evolutionary link to Ser/Thr protein phosphatases?
BMC Res Notes. 1: 78. 19. Zhao, A. et al (2011) Rapid isolation of high-affinity human antibodies against the tumor vascular marker Endosialin/TEM1, using a paired yeast-display/secretory scFv library platform.
J Immunol Methods. 363: 221-32. 20. Patino, G.A. et al. (2011) Voltage-Gated Na+ Channel {beta}1B: A Secreted Cell Adhesion Molecule Involved in Human Epilepsy.
J Neurosci. 31: 14577-91. 21. Gatherer, D. et al. (2011) High-resolution human cytomegalovirus transcriptome.
Proc Natl Acad Sci U S A.108: 19755-60. 22. Mahuzier, A. et al. (2012) Dishevelled stabilization by the ciliopathy protein Rpgrip1l is essential for planar cell polarity.
J Cell Biol. 198: 927-40. 23. Zhao, C. et al. (2005) Human ISG15 conjugation targets both IFN-induced and constitutively expressed proteins functioning in diverse cellular pathways.
Proc Natl Acad Sci U S A. 102:10200-5 24. Singh, A. et al. (2014) Trypanosome MKT1 and the RNA-binding protein ZC3H11: interactions and potential roles in post-transcriptional regulatory networks.
Nucleic Acids Res. 42: 4652-68. 25. Mui MZ et al. (2015) The human adenovirus type 5 E4orf4 protein targets two phosphatase regulators of the Hippo signaling pathway.
J Virol. pii: JVI.03710-14.

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